Description

The Q5® High-Fidelity 2X Master
Mix offers robust, high-fidelity performance in a
convenient master mix format. The Q5 High-
Fidelity 2X Master Mix features a high-fidelity,
thermostable DNA polymerase with 3´→ 5´
exonuclease activity, fused to a processivity-enhancing
Sso7d domain to support robust DNA
amplification. With an error rate > 100-fold lower
than that of Taq DNA Polymerase and 12-fold
lower than that of Pyrococcus furiosus (Pfu) DNA
Polymerase, Q5 High-Fidelity DNA Polymerase
is ideal for cloning and can be used for long or
difficult amplicons. The convenient master mix
formulation is supplied at a 2X concentration.
The mix contains dNTPs, Mg++ and a proprietary
broad-use buffer requiring only the addition of
primers and DNA template for robust amplification
regardless of GC content. When used at the
recommended 1X final concentration, the Q5
High-Fidelity Master Mix contains 2 mM MgCl2.
Q5 High-Fidelity DNA Polymerase is unlike typical,
lower fidelity PCR enzymes. To determine the optimal annealing temperatures for a given set of primers, use of the NEB Tm Calculator is highly recommended.

Q5-High Fidelity 2X Master Mix formats allow robust amplification of a broad range of targets with a single formulation. Amplification of a variety of human genomic amplicons from
low to high GC content using Q5 High-Fidelity 2X Master Mix. All reactions were
conducted using 30 cycles of amplification and visualized by microfluidic
LabChip® analysis.

Notes

A precipitate (most noticeable after the first 1–2 freeze/thaw cycles) is not uncommon. To ensure optimal performance, the master mix should be thawed and resuspended prior to use. Stability testing using up to 15 freeze/thaw cycles has shown no negative effect on master mix performance.

Product specifications for individual components in the Q5 High-Fidelity 2X Master Mix are available separately.

Protocols

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Usage Guidelines & Tips

Troubleshooting Guides

Application Notes

Quality Control

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.

PCR Amplification (Master Mix):The polymerase master mix is tested in a polymerase chain reaction (PCR) using a control template and specific primers, resulting in the expected product.

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Legal and Disclaimers

Licenses

This product is licensed from Bio-Rad Laboratories, Inc. under U.S. Pat. Nos.
6,627,424, 7,541,170, 7,670,808, 7,666,645 and corresponding patents in other
countries for use only in: (a) standard (non-real time) PCR in the research
field only, but not real-time PCR or digital PCR; (b) any in-vitro
diagnostics application, except for applications using real-time or digital PCR;
and (c) any non-PCR applications in DNA sequencing, isothermal amplification and
the production of synthetic DNA.